title: Analysis of histones in Xenopus laevis. II. mass spectrometry reveals an index of cell type-specific modifications on H3 and H4.
abstract: Epigenetic information is hypothesized to be encoded in histone variants and post-translational modifications. Varied cell- and locus-specific combinations of these epigenetic marks are likely contributors to regulation of chromatin-templated transactions, including transcription, replication, recombination, and repair. Therefore, the relative abundance of histone modifications in a given cell type is a potential index of cell fate and specificity. Here, we utilize mass spectrometry techniques to characterize the relative abundance index of cell type-specific modifications on histones H3 and H4 in distinct cell types from the frog Xenopus laevis, including the sperm, the stored predeposition histones in the egg, the early embryo equivalent pronuclei, cultured somatic cells, and erythrocytes. We used collisionally associated dissociation to identify the modifications present on histone H3 in a variety of cell types, resolving 26 distinctly modified H3 peptides. We employed the electron transfer dissociation fragmentation technique in a "middle-down" approach on the H4 N-terminal tail to explore the overlap of post-translational modifications. We observed 66 discrete isoforms of the H4 1-23 fragment in four different cell types. Isolation of the stored, predeposition histone H4 from the frog egg also revealed a more varied pattern of modifications than the previously known diacetylation on Lys(5) and Lys(12). The developmental transitions of modifications on H3 and H4 were strikingly varied, implying a strong correlation of the histone code with cell type and fate. Our results are consistent with a histone code index for each cell type and uncover potential cross-talk between modifications on a single tail.
authors.affiliation: Laboratory of Chromatin Biology, The Rockefeller University, New York, New York 10065, USA.
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: Nicklay
authors.forename: David
authors.initials: D
authors.lastname: Shechter
authors.forename: Raghu K
authors.initials: RK
authors.lastname: Chitta
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Garcia
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Shabanowitz
authors.forename: C David
authors.initials: CD
authors.lastname: Allis
authors.forename: Donald F
authors.initials: DF
authors.lastname: Hunt
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D006657:Histones / Q000032:analysis* / Q000737:chemistry* / Q000145:classification / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D013379:Substrate Specificity; D014982:Xenopus laevis / Q000378:metabolism
chemical_list: D006657:Histones